Evaluation of the impact of ul54 gene-deletion on the global transcription and DNA replication of pseudorabies virus

Abstract

Pseudorabies virus (PRV) is an animal alpha-herpesvirus with a wide host range. PRV has 67 protein-coding genes and several non-coding RNA molecules, which can be classified into three temporal groups, immediate early, early and late classes. The ul54 gene of PRV and its homolog icp27 of herpes simplex virus have a multitude of functions, including the regulation of viral DNA synthesis and the control of the gene expression. Therefore, abrogation of PRV ul54 function was expected to exert a significant effect on the global transcriptome and on DNA replication. Real-time PCR and real-time RT-PCR platforms were used to investigate these presumed effects. Our analyses revealed a drastic impact of the ul54 mutation on the genome-wide expression of PRV genes, especially on the transcription of the true late genes. A more than two hour delay was observed in the onset of DNA replication, and the amount of synthesized DNA molecules was significantly decreased in comparison to the wild-type virus. Furthermore, in this work, we were able to successfully demonstrate the utility of long-read SMRT sequencing for genotyping of mutant viruses.

Kyllinga nemoralis Methanolic Roots Extract Inhibits Herpes Simplex Virus Type 1 Replication Cycle

“…Besides that, the UL54 gene affects the functions of other early and late genes, including gC for viral attachment. 20 It encodes DNA polymerase catalytic enzyme that important proteins during the viral lytic cycle and increasing the transcription of the late genes. 21,22 Based on the findings, at the immediate early replication phase (2 hpi), the K. nemoralis methanolic extract caused all three genes to be down-regulated because of the alteration and disordering of cell receptors and viral glycoproteins during virus attachment and entry.…”

Resveratrol Inhibits Pseudorabies Virus Replication by Targeting IE180 Protein

“…The IE180, the only IE gene of PRV, is a transcription factor and is considered to be essential for the transcription of all known genes ( Tombácz et al, 2017 ). IE180 is highly similar to the IE proteins of other Alphaherpesviruses such as ICP4 of HSV-1, IE140 of VZV, IE1 of equine herpesvirus1, and p180 of bovine herpesvirus1 ( Csabai et al, 2017 ). All herpesviruses have the same regulatory mechanism.…”

Analysis of the Complete Genome Sequence of a Novel, Pseudorabies Virus Strain Isolated in Southeast Europe

“…For the isolation of virions, PK-15 cells were infected with PRV-MdBio using MOI = 0.1 pfu/cell and then incubated until complete cytopathic effect was observed. Virions were separated from the PK-15 cells by ultracentrifugation (Sorwall WX Ultra 90, Thermo Scientific) on a 30% sucrose cushion as has been described earlier [46]. The viral DNA was extracted from the samples using the traditional phenol/chloroform method.…”